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Ultrastructural evidence for multiple mucous domains in frog olfactory epithelium

Summary

This study showed that the olfactory mucus is a highly structured extracellular matrix. Several olfactory epithelial glycoconjugates in the frog Rana pipiens were localized ultrastructurally using rapid-freeze, freeze-substitution and post-embedding (Lowicryl K11M) immunocytochemistry. Two of these conjugates were obtained from membrane preparations of olfactory cilia, the glycoproteins gp95 and olfactomedin. The other conjugates have a carbohydrate group which in the olfactory bulb appears to be mostly on neural cell-adhesion molecules (N-CAMs); in the olfactory epithelium this carbohydrate is present on more molecules. Localization of the latter conjugates was determined with monoclonal antibodies 9-OE and 5-OE. Ultrastructurally all antigens localized in secretory granules of apical regions of frog olfactory supporting cells and in the mucus overlying the epithelial surface, where they all had different, but partly overlapping, distributions. Monoclonal antibody 18.1, to gp95, labeled the mucus throughout, whereas poly- and monoclonal anti-olfactomedin labeled a deep mucous layer surrounding dendritic endings, proximal parts of cilia, and supporting cell microvilli. Labeling was absent in the superficial mucous layer, which contained the distal parts of the olfactory cilia. Monoclonal antibody 9-OE labeled rather distinct areas of mucus. These areas sometimes surrounded dendritic endings and olfactory cilia. Monoclonal antibody 5-OE labeled membranes of dendritic endings and cilia, and their glycocalyces, and also dendritic membranes.

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References

  1. Becker A, Peters W (1989) Localization of sugar-binding sites in contact chemosensilla of Periplaneta americana. J Insect Physiol 35:239–250

  2. Bendayan M, Garzon S (1988) Protein G-gold complex: comparative evaluation with protein A-gold for high-resolution immunocytochemistry. J Histochem Cytochem 36:597–607

  3. Burton PR (1985) Ultrastructure of the olfactory neuron of the bullfrog: the dendrite and its microtubules. J Comp Neurol 242:147–160

  4. Burton PR (1992) Ultrastructural studies of the microtubules and microtubule-organizing centers of the vertebrate olfactory neuron. Microsc Res Techn 23: in press

  5. Carlemalm E, Villiger W (1989) Low temperature embedding. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 4. Academic Press, London, pp 29–45

  6. Chen Z, Ophir D, Lancet D (1986a) Monoclonal antibodies to ciliary glycoproteins of frog olfactory neurons. Brain Res 368:329–338

  7. Chen Z, Pace U, Heldman J, Shapira A, Lancet D (1986b) Isolated frog olfactory cilia: a preparation of dendritic membranes from chemosensory neurons. J Neurosci 6:2146–2154

  8. Chen Z, Pace U, Ronen D, Lancet D (1986c) Polypeptide gp95. A unique glycoprotein of olfactory cilia with transmembrane receptor properties. J Biol Chem 261:1299–1305

  9. Cuschieri A, Bannister LH (1974) Some histochemical observations on the mucosubstances of the nasal glands of the mouse. Histochem J 6:543–558

  10. Dear TN, Boehm T, Keverne EB, Rabbits TH (1991) Novel genes for potential ligand-binding proteins in subregions of the olfactory mucosa. EMBO J 10:2813–2819

  11. Dear TN, Campbell K, Rabbits TH (1991) Molecular cloning of putative odorant-binding and odorant-metabolizing proteins. Biochemistry 30:10376–10382

  12. Foster JD, Getchell ML, Getchell TV (1991) Identification of sugar residues in secretory glycoconjugates of olfactory mucosae by using lectin histochemistry. Anat Rec 229:525–544

  13. Foster JD, Getchell ML, Getchell TV (1992) Ultrastructural localization of sialylated glycoconjugates in cells of the salamander olfactory mucosa using lectin cytochemistry. Cell Tissue Res 267:113–124

  14. Getchell ML, Getchell TV (1991) Immunohistochemical localization of components of the immune barrier in the olfactory mucosae of salamanders and rats. Anat Rec 231:358–374

  15. Getchell ML, Getchell TV (1992) Fine structural aspects of secretion and extrinsic innervation in the olfactory mucosa. Microsc Res Techn 23: in press

  16. Getchell ML, Mellert TK (1991) Olfactory mucus secretion. In: Getchell TV, Doty RL, Bartoshuk LM, Snow JB (eds) Smell and taste in health and disease. Raven Press, New York, pp 83–95

  17. Getchell ML, Rafols JA, Getchell TV (1984) Histological and histochemical studies of the secretory components of the salamander olfactory mucosa: effects of isoproterenol and olfactory nerve section. Anat Rec 208:553–565

  18. Getchell ML, Zielinski B, DeSimone JA, Getchell TV (1987) Odorant stimulation of secretory and neural processes in the salamander olfactory mucosa. J Comp Physiol [A] 160:155–168

  19. Getchell ML, Zielinski B, Getchell TV (1988) Odorant and autonomic regulation of secretion in the olfactory mucosa. In: Margolis FL, Getchell TV (eds) Molecular neurobiology of the olfactory system. Molecular, membranous, and cytological studies. Plenum Press, New York, pp 71–98

  20. Getchell ML, Bouvet J-F, Finger TE, Holley A, Getchell TV (1989) Peptidergic regulation of secretory activity in amphibian olfactory mucosa: immunohistochemistry, neural stimulation, and pharmacology. Cell Tissue Res 256:381–389

  21. Getchell TV, Margolis FL, Getchell ML (1984) Perireceptor and receptor events in vertebrate olfaction. Progr Neurobiol 23:317–345

  22. Gladysheva O, Kukushkina D, Martynova G (1986) Glycoprotein composition of olfactory mucus in vertebrates. Acta Histochem (Jena) 78:141–146

  23. Key B, Akeson RA (1990a) Olfactory neurons express a unique glycosylated form of the neural cell adhesion molecule (N-CAM). J Cell Biol 110:1729–1743

  24. Key B, Akeson RA (1990b) Immunochemical markers for the frog olfactory neuroepihelium. Dev Brain Res 57:103–117

  25. Key B, Akeson RA (1991) Delineation of olfactory pathways in the frog nervous system by unique glycoconjugates and N-CAM isoforms. Neuron 6:381–396

  26. Kinnamon SC, Getchell TV (1991) Sensory transduction in olfactory receptor neurons and gustatory receptor cells. In: Getchell TV, Doty RL, Bartoshuk LM, Snow JB (eds) Smell and taste in health and disease. Raven Press, New York, pp 145–172

  27. Leunissen JLM (1990) Background suppression using Aurison BSA-C and/or Tween-20®. Aurion Newsletter 1 (Wageningen, The Netherlands) October 1990

  28. Mair RG, Bouffard JA, Engen T, Morton TH (1978) Olfactory sensivity during the menstrual cycle. Sens Proc 2:90–98

  29. Menco BPM (1980) Qualitative and quantitative freeze-fracture studies on olfactory and nasal respiratory structures of frog, ox, rat, and dog. I. A general survey. Cell Tissue Res 207:183–209

  30. Menco BPM (1989) Olfactory and nasal respiratory epithelia, and foliate taste buds visualized with rapid-freeze freeze-substitution and Lowicryl K11M embedding. Ultrastructural and initial cytochemical studies. Scanning Microsc 3:257–272

  31. Menco BPM (1991) Ultrastructural localization of antibodies to olfactory epithelial-specific glycoproteins gp95, olfactomedin, and 9-OE and 5-OE antigens in the frog's olfactory epithelium (abstract). Chem Senses 16:554–555

  32. Menco BPM (1992a) Ultrastructural studies on membrane, cytoskeletal, mucous and protective compartments in olfaction. Microsc Res Techn 22:215–224

  33. Menco BPM (1992b) Lectins bind differentially to cilia and microvilli of major and minor cell populations in olfactory and nasal respiratory epithelia. Microsc Res Techn (in press)

  34. Menco BPM, Bruch RC, Dau B, Danho W (1992) Ultrastructural localization of olfactory transduction components: the G-protein subunit Golfα and type III adenylyl cyclase. Neuron 8:441–453

  35. Menco BPM, Hellekant G (1993) Ultrastructural evidence for a binding substance to the sweet-tasting protein thaumatin inside taste bud pores of rhesus monkey foliate papillae. Microsc Res Techn (in press)

  36. Okano M, Takagi SF (1974) Secretion and electrogenesis of the supporting cell in the olfactory epithelium. J Physiol (Lond) 242:353–370

  37. Phillips GW, Bridgman PC (1991) Immunoelectron microscopy of acetylcholine receptors and 43 kD protein after rapid freezing, freeze-substitution, and low-temperature embedding in Lowicryl K11M. J Histochem Cytochem 39:625–634

  38. Phillips TE, Boyne AF (1984) Liquid nitrogen-based quick freezing: experiences with bounce-free delivery of cholinergic nerve terminals to a metal surface. J Electron Microsc Tech 1:9–29

  39. Proctor DF (1982) The mucociliary system. In: Proctor DF, Andersen IB (eds) The nose: upper airway physiology and the admospheric environment. Elsevier, Amsterdam, pp 245–278

  40. Puchelle É, Beorchia A, Ménager M, Zahm J-M, Ploton D (1991) Three-dimensional imaging of the mucus secretory process in the cryofixed frog respiratory epithelium. Biol Cell: 159–166

  41. Reese TS (1965) Olfactory cilia in the frog. J Cell Biol 25:209–230

  42. Sbarbati A, Ceresi E, Accordini C (1991) Surfactant-like material on the chemoreceptorial surface of the frog's taste organ: an ultrastructural and electron spectroscopic imaging study. J Struct Biol 107:128–135

  43. Snyder DA, Rivers AM, Yokoe H, Menco BPM, Anholt RRH (1991) Olfactomedin: purification, characterization and localization of a novel olfactory glycoprotein. Biochemistry 30:9143–9153

  44. Steinbrecht RA (1992) Fine structure of insect olfactory receptor cell complexes. Microsc Res Techn 22:336–350

  45. Strous GJ, Dekker J (1992) Mucin-type glycoproteins. Crit Rev Biochem Mol Biol 27:57–92

  46. Sturgess JM (1979) Mucous secretions in the respiratory tract. Pediatr Clin North Am 26:481–501

  47. Takagi SF (1989) Human olfaction. University of Tokyo Press

  48. Takagi SF, Okano M (1974) An electrophysiological and electron-microscopical study of the secretion in the olfactory epithelium. Gunma Symp Endocr 11:3–14

  49. Yamamoto T, Tonosaki A, Kurosawa T (1965) Electron microscope studies on the olfactory epithelium of frogs. Acta Anat Nippon 40:342–353

  50. Zielinski BS, Getchell ML, Getchell TV (1988) Ultrastructural characteristics of sustentacular cells in control and odoranttreated olfactory mucosae of the salamander. Anat Rec 221:769–779

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Correspondence to Bert P. M. Menco.

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Menco, B.P.M., Farbman, A.I. Ultrastructural evidence for multiple mucous domains in frog olfactory epithelium. Cell Tissue Res. 270, 47–56 (1992). https://doi.org/10.1007/BF00381878

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Key words

  • Cilia
  • Microvilli
  • Secretory granules
  • Mucus
  • Freeze-substitution
  • Immunocytochemistry
  • Rana pipiens (Anura)